摘要
目的:探讨miR-24对肝癌MHCC97H细胞增殖、凋亡及迁移能力的影响及其可能机制。方法:分别构建miR-24及其反义序列(anti-miR-24)的高表达质粒,分别转染肝癌MHCC97H细胞,根据质粒转染情况将实验细胞分为空白对照组、anti-miR-24组和miR-24高表达组。应用CCK-8法检测肝癌MHCC97H细胞的增殖能力;流式细胞仪检测细胞的凋亡率;划痕实验观察细胞的迁移能力;逆转录—聚合酶链反应(RT-PCR)检测Bcl-2的mRNA表达水平,免疫组织化学法和western blotting法检测Bcl-2的蛋白表达水平。结果:与空白对照组相比,miR-24高表达组肝癌MHCC97H细胞增殖和迁移能力明显降低(P<0.05),而anti-miR-24组细胞增殖和迁移能力升高(P<0.05);miR-24高表达组细胞凋亡率较空白对照组升高(P<0.05),anti-miR-24组的细胞凋亡率较空白对照组降低(P<0.05)。与对照组相比,miR-24高表达组Bcl-2mRNA及其蛋白表达水平均降低(P<0.05),而anti-miR-24组Bcl-2mRMA及其蛋白表达水平显著上升(P<0.05)。结论:miR-24可能通过下调Bcl-2的表达,促进肝癌MHCC97H细胞凋亡。
Objective:To investigate the effect of miR-24 on the proliferation,apoptosis and migration of MHCC97H and to explore the underlying mechanism.Methods:The miR-24 and anti-miR-24 high expression plasmid were constructed.The MHCC97H were transfected with miR-24 and anti-miR-24 high expression plasmid,and than were divided into the blank control group,anti-miR-24 group and miR-24 group according to transfection.The proliferation ability of MHCC97H was detected by CCK8,the cells apoptosis rate was measured by flow cytometry,the migration capacity of MHCC97H was detected by cell scratch test.RT-PCR was used to detect the expression of Bcl-2 mRNA.Immunocytochemistry and western blotting were used to detect the expression of Bcl-2 protein in MHCC97H.Results:The proliferation and migration capacity of miR-24 high expression plasmid group was obviously decreased compared with blank control group(P<0.05),but the proliferation and migration ability in anti-miR-24 gruop was significantly increased(P<0.05).Compared with the blank control group,the apoptosis rate of miR-24 high expression plasmid group was increased(P<0.05),while it was decreased in anti-miR-24 group(P<0.05).The expression level of Bcl-2 mRNA and protein was decreased after over-expression of miR-24(P<0.05).Conclusion:miR-24 might promote the apoptosis of liver cancer MHCC97H cells by the down-regulation of Bcl-2 expression.
作者
杨鹏
滕红丽
罗雪兰
李丹
颜鸳渊
欧和生
覃裕旺
Yang Peng;Teng Hongli;Luo Xuelan;Li Dan;Yan Yuanyuan;Ou Hesheng;Qin Yuwang(Guangxi Internationnal Zhuang Medicine Hospital,Nanning 530201,China;College of Pharmacy,Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2019年第9期1397-1402,共6页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.81373403)
广西创新驱动发展专项资金资助项目(No.桂科AA17202034)
广西壮族自治区教育厅中青年教师基础能力提升课题资助项目(No.2019KY0344)
